CD4+ T-Cell Dysfunction in Severe COVID-19 Disease Is Tumor Necrosis Factor-α/Tumor Necrosis Factor Receptor 1-Dependent.

Rationale: Lymphopenia is common in severe coronavirus disease (COVID-19), yet the immune mechanisms are poorly understood. As inflammatory cytokines are increased in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, we hypothesized a role in contributing to reduced T-cell numbers. Objectives: We sought to characterize the functional SARS-CoV-2 T-cell responses in patients with severe versus recovered, mild COVID-19 to determine whether differences were detectable. Methods: Using flow cytometry and single-cell RNA sequence analyses, we assessed SARS-CoV-2-specific responses in our cohort. Measurements and Main Results: In 148 patients with severe COVID-19, we found lymphopenia was associated with worse survival. CD4+ lymphopenia predominated, with lower CD4+/CD8+ ratios in severe COVID-19 compared with patients with mild disease (P < 0.0001). In severe disease, immunodominant CD4+ T-cell responses to Spike-1 (S1) produced increased in vitro TNF-α (tumor necrosis factor-α) but demonstrated impaired S1-specific proliferation and increased susceptibility to activation-induced cell death after antigen exposure. CD4+TNF-α+ T-cell responses inversely correlated with absolute CD4+ counts from patients with severe COVID-19 (n = 76; R = -0.797; P < 0.0001). In vitro TNF-α blockade, including infliximab or anti-TNF receptor 1 antibodies, strikingly rescued S1-specific CD4+ T-cell proliferation and abrogated S1-specific activation-induced cell death in peripheral blood mononuclear cells from patients with severe COVID-19 (P < 0.001). Single-cell RNA sequencing demonstrated marked downregulation of type-1 cytokines and NFκB signaling in S1-stimulated CD4+ cells with infliximab treatment. We also evaluated BAL and lung explant CD4+ T cells recovered from patients with severe COVID-19 and observed that lung T cells produced higher TNF-α compared with peripheral blood mononuclear cells. Conclusions: Together, our findings show CD4+ dysfunction in severe COVID-19 is TNF-α/TNF receptor 1-dependent through immune mechanisms that may contribute to lymphopenia. TNF-α blockade may be beneficial in severe COVID-19.

Chitosan functionalized Halloysite Nanotubes as a receptive surface for laccase and copper to perform degradation of chlorpyrifos in aqueous environment.

Chitosan (CTS) functionalized Halloysite Nanotubes (HNT) have been used as receptive nano-supports for the grafting of copper (Cu) and laccase (Lac) for the degradation of chlorpyrifos. The developed nanocomposite Lac@Cu-CTS-HNT showed 83.4% Lac immobilization which was further characterized by TEM, SEM-EDX, FTIR, XRD, DSC and TGA. The chlorpyrifos degradation studies were performed under constant stirring for 24 h with both free enzyme and Lac@Cu-CTS-HNT and were analysed through HPLC. Percentage degradation of chlorpyrifos with the nanocomposite went as high as 97% for 50 µg/mL chlorpyrifos at neutral pH and room temperature. Variable pesticide and nanocomposite concentration, pH, and temperature studies for pesticide degradation were also performed, followed by reusability studies. The nanocomposite maintained its degradation ability at ~97% even at variable temperature and pH conditions. Reusability study was performed 5 times wherein the degradation percentage remained the same after 5 cycles (~<95%). Degradation kinetics were also performed for the nanocomposite in the presence and absence of the immobilized enzyme. Through this study, it is suggested that Lac@Cu-CTS-HNT can be a potential nano-catalyst for the degradation of chlorpyrifos in aqueous environment.